This study was aimed to investigate the physicochemical changes induced in 200 nm extruded oligolamellar DPPC:DPPC (10:1) liposomes by freezing, followed by gamma-irradiation, in the absence and presence of 5 mM stable cyclic nitroxide radicals, 2,2,6,6-tetramethylpiperidine-1-oxyl (Tempo) and 4-hydroxy-2,2,6, 6-tetramethylpiperidine-1-oxyl (Tempol). The char acterization is based on the use of differential scanning calorimetry (DSC) and was aimed to differentiate the contribution of freezing and gamma-irradiation in the presence and absence of nitroxides. Liposomal preparations of DPPC/DPPG which have sub-, pre- and main-phase transitions in the temperature range (0 degrees C < T-m < 50 degrees C) were used. Our results show that: (1) freezing modified and induced fusion to MLV as well as fission to SUV, (2) freezing did not fully prevent the radiation-induced changes in the thermotropic characteristics of the liposomes, and (3) Tempo and Tempol did not prevent the changes in thermotropic behavior caused as a result of freezing of the liposomal dispersion. These results demonstrate that DSC is a powerful and sensitive tool in both physical and chemical studies of lipid assemblies.